THE COMPLEMENT SYSTEM (HUMORAL AND CHEMICAL (endogenous) BARRIERS.

OUTLINE: Definition and brief historical introduction Classes of complement pathways Function or main roles Mechanism of action Protection of host cells and some diseases

COMPLEMENT SYSTEM: DEFINITION: Is a system of interacting proteins in the blood that can be activated by antigenantibody reaction or micro-organisms to undergo a cascade of proteolytic reactions whose end result is the assembly of membrane attack complexes (MAC). The MAC helps to kill invading pathogens or cells and as such defend the body against such. HISTORICAL OVERVIEW: In the late 19th century, blood serum was found to contain a "factor" or "principle" capable of killing bacteria. In 1896, Jules Bordet, a young Belgian scientist in Paris at the Pasteur Institute, demonstrated that this principle could be analyzed into two components: a heat-stable and a heat-labile component. (Heat-labile means that the component loses its effectiveness if the serum is heated.) The heat-stable component was found to confer immunity against specific microorganisms, whereas the heat-labile component was found to be responsible for the non-specific antimicrobial activity conferred by all normal serum. This heat-labile component is what we now call "complement." The term "complement" was introduced by Paul Ehrlich in the late 1890s, as part of his larger theory of the immune system. According to this theory, the immune system consists of cells that have specific receptors on their surface to recognize antigens. Upon immunization with an antigen, more of these receptors are formed, and they are then shed from the cells to circulate in the blood. These receptors, which we now call "antibodies," were called by Ehrlich "amboceptors" to emphasize their bifunctional binding capacity: They recognize and bind to a specific antigen, but they also recognize and bind to the heat-labile antimicrobial component of fresh serum. Ehrlich, therefore, named this heat-labile component "complement," because it is something in the blood that "complements" the cells of the immune system

The rational for the inclusion of complements in the innate immune response is that: i. ii. It is not adaptable It does not change over the course of ones lifetime.

However, it can be recruited and brought into action by the adaptive immune system VARIOUS CLASSES OF THE COMPLEMENT PATHWAY: Three (3) main biochemical pathways activate complement system 1. The classical complement pathway 2. The alternative complement pathway 3. Mannose binding lectin pathway

The following are the basic functions of the complement 1. Lysis of cells, bacteria and viruses. 2. Opsonization, which promotes phagocytosis of particulate antigens. 3. Binding to specific complement receptors on the cells of the immune system, triggering specific cell functions, inflammation, and certain immunoregulatory molecules. 4. Immune Clearance, which removes immune complexes from immune system and deposits them in the spleen and liver. GENERAL OVERVIEW OF MECHANISM OF ACTION The complement system has about 20 different proteins circulating as inactive zymogens. These are produced by different cells including i. ii. iii. Hepatocytes Macrophages Gut epithelial cells

The majority is produced by the hepatocytes. The reacting components of the complement system are designated C1 to C9, Factor B and Factor D. The rest are kind of regulatory proteins. The protein components in the complement system are all soluble proteins.

In terms or function, complement proteins are subdivided into 3. These are:

C1 C4, C2, C3 C5 to C9

RECOGNITION ACTIVATION ATTACK

Another way of classification employs the time phase of the whole process. Hence we have the EARLY PHASE, THE INTERMEDIATE PHASE & LATE PHASE.

The main role of the complement is to attack the membrane of microbial cells therefore activation of complement is focused on the microbial cell membrane where it is triggered either by antibody bound to the microorganism or by microbial envelope polysaccharide both of which activate the early complement.

There are 2 sets of the early complements belonging to the 2 distinct pathways.

C1, C2, C4 Factor B and Factor D

Classical Pathway Alternative pathway

Triggered by antibody Triggered by microbial polysaccharides

The early components act on C3 the most important complement component. These early components are pro-enzymes (C1, C4, C2) activated sequentially by limited proteolytic cleavage. As each pro-enzyme in the sequence is cleaved, it is activated to generate a serine protease which cleaves the next pro-enzyme in the sequence. Many of the cleavages liberate 1. A small peptide fragment 2. A large fragment which has an exposed membrane binding site on it. Larger fragments bind tightly to the target cell membrane by its newly exposed membrane binding site and helps to carry out the next reaction in the sequence. In this way complement activation is confined largely to the cell surface where it begun. The smaller fragments act independently as a diffusible signal that promotes an inflammatory response. C3 activation is the central reaction in the complement activation sequence and it is here that the classical and alternative pathways converge. For both pathways, C3 is cleaved by an enzyme complex called C3 convertase though different convertase is produced by each pathway. Convertase are formed by the spontaneous assembly of 2 of the complement component activated earlier in the cascade. Both cleave C3 into two fragments. The larger C3b binds covalently to the target cell membrane and then binds to C5. Once bound, the C5 protein is cleaved by the C3 convertase now acting as C5 convertase to initiate the spontaneous assembly of the late component.

THE ALTERNATIVE PATHWAY Some polysaccharides in the cell envelope of microorganisms can activate the alternative pathway. This pathway provides the first line of defense against infection before an immune response can be mounted and it amplifies the effect of the classical once an immune response has begun. C3 is spontaneously hydrolysed to C3a and C3b due to the breakdown of the thio-ester bond through condensation reaction. The C3b binds to the pathogenic membrane surface if pathogen is near enough. If there is no pathogen in the blood C3a and C3b protein fragments will be deactivated by rejoining with each other. When C3b binds to the pathogenic cell membrane, Factor B binds to the membrane bound C3b to form C3bB. Factor D which circulates in the blood in an active form cleaves the bound factor B to generate the active fragment Bb and therefore produce C3bBb (which is the C3 convertase for the alternative pathway) and therefore generates more C3b molecules some of which binds C5. The C3 convertase can also act as a C5 convertase and cleave the membrane bound C5 molecules to initiate the assembly of the Membrane Attack Complex which is involves the C5b bound to the C3b binding to the C6 to form C56 and then C7 to form C567. The C567 complex then binds firmly via the C7 to the membrane. This complex adds one molecule of C8 to form C5678 which then binds 8 to 18 molecules of C9 which partially unfold and polymerise into a transmembrane channel. The Membrane Attack Complexes form aqueous pores through the membrane this compromise the structure of the lipid bilayer in their vicinity and they make the membrane leaky. Small molecules leak in and out of the cell around and through the complexes while macromolecules remain inside. The cells normal mechanism for controlling water balance is disrupted. Water is therefore drawn into the cell by osmosis causing it to swell and burst. HOST CELL PROTECTION The complement system has the potential to be extremely damaging to host tissues, meaning its activation must be tightly regulated. The complement system is regulated by complement control proteins, which are present at a higher concentration in the blood plasma than the complement proteins themselves. Some complement control proteins are present on the membranes of selfcells preventing them from being targeted by complement. One example is CD59, also known as protectin which inhibits C9 polymerisation during the formation of the membrane attack complex.

SOME DISEASES ASSOCIATED WITH THE COMPLEMENT SYSTEM It is thought that the complement system might play a role in many diseases with an immune component, such as Barraquer-Simons Syndrome, asthma, lupus erythematosus, glomerulonephritis, various forms of arthritis, autoimmune heart disease, multiple sclerosis, inflammatory bowel disease, and ischemia-reperfusion injuries. The complement system is also becoming increasingly implicated in diseases of the central nervous system such as Alzheimer's disease and other neurodegenerative conditions. Deficiencies of the terminal pathway predispose to both autoimmune disease and infections (particularly Neisseria meningitis, due to the role that the C56789 complex plays in attacking Gram-negative bacteria

References Molecular Biology of the Cell, 2nd Edition, Bruce Alberts, Dennis Bray, Julius Lewis et al, Published by Garland Publishers 1999, Page 1032-1033 Human Physiology, 5th Edition, S I Fox, Published by McGraw Hill Companies Inc, 1996 www.wikipedia.com